Penicillins

ABSTRACT

New penicillins and their salts and esters particularly active against Gram-negative organisms such as Pseudomonas spp., and their preparation and administration. The penicillins are active against clinically important organisms against which well-known broad spectrum penicillins are inactive and may generally be designated as aminoacyldipeptide penicillins of unusual structure and properties.

CROSS-REFERENCE

This is a division of Serial No. 466,814 filed May 3, 1974.

This invention relates to penicillins which have, in general, a broad spectrum of antibacterial activity, being active against many species of Gram-positive and Gram-negative bacteria. They are thus useful as therapeutic (and, to a lesser extent, prophylactic) agents in animals, including man and poultry. The invention further relates to methods for the preparation of these penicillins and to their use in therapy.

Although there are now available a number of semi-synthetic penicillins having what is known as broad-spectrum activity, no single penicillin is yet available which has a clinically useful level of antibacterial activity against all the pathogenic organisms encountered in clinical practice. The search thus continues for broad-spectrum penicillins which have advantages, either in improved antibacterial effectiveness or wider spectrum of activity, over the available penicillins.

According to the present invention there is provided a penicillin of formula (I) or a pharmaceutically acceptable salt or ester thereof: ##SPC1##

Wherein

R is pnenyl, phenyl substituted by one or more functional groups selected from hydroxy, halogen, nitro, alkoxy containing from 1 to 3 carbon atoms, and amino groups, 2- or 3 -thienyl, cycloalkyl having from 3 to 7 carbon atoms, cycloalkenyl having from 5 to 7 carbon atoms or alkyl having from 1 to 4 carbon atoms;

R³ is hydrogen or an alkyl group having from 1 to 3 carbon atoms; R¹ is hydrogen or an organic radical containing up to 20 carbon atoms;

R² is a group of formula (II) or (III): ##EQU1## wherein R⁵ is amino, mono-or di-alkylamino wherein the alkyl groups contain from 1 to 4 carbon atoms, cyclohexylamino, hydrogen, alkyl having from 1 to 4 carbon atoms, or phenyl and R⁶ is amino or mono- or di-alkylamino wherein the alkyl groups contain from 1 to 4 carbon atoms, or cyclohexylamino.

The group R may be, for example, phenyl, 4-hydroxyphenyl, 3-chloro-4-hydroxyphenyl, 4-nitrophenyl, 4-chlorophenyl, 4-fluorophenyl, 4-methoxy-phenyl, 4-aminophenyl, 2-thienyl, 3-thienyl, cyclopropyl, cyclohexyl, cyclohexa-1,4-dienyl, iso-propyl or methyl.

The group R¹ may, for example, be hydrogen, phenyl, 4-hydroxyphenyl, 4-nitrophenyl, 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, 4-aminophenyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, methylene, ethylene, ethylthio, n-propoxy-methyl, carbamyl, carbamylmethyl, acetoxy, phenoxy, benzyloxy, 2-thienyl, 3-thienyl, indol-3-yl, 1H-imidazol-5-yl, cyclohexa-1,4-dienyl, cyclopropyl or cyclohexyl.

The group R⁵ may, for example, be amino, methylamino, n-butylamino, tert-butylamino, cyclohexylamino, hydrogen, methyl, ethyl, n- or isopropyl, n-, sec, or tert-butyl, or phenyl.

The group R⁶ may for example, be amino, methylamino, dimethylamino, ethylamino, isopropylamino, n-propylamino, iso-propylamino, tert-butylamino, n-butylamino or cyclohexylamino.

Preferably R is phenyl, 4-hydroxyphenyl, or 3-thienyl.

Preferably R¹ is phenyl, 4-hydroxyphenyl, 4-chlorophenyl, 4-fluorophenyl, 4-nitrophenyl, 3-indolyl, or methylthiomethyl.

Preferably R³ is hydrogen.

Preferably R⁵ is amino or hydrogen.

Preferably R⁶ is amino.

Preferably the carbon atom to which the group R in formula (I) is attached is in the D configuration.

Preferably the carbon atom to which the group R² in formula (I) is attached is in the D configuration.

Examples of suitable salts of compounds (I) include the sodium, potassium, calcium, magnesium or aluminium salts, and ammonium or substituted ammonium salts, for example those with trialkylamines such as triethylamine, procaine, dibenzylamine, triethanolamine, 1-ephenamine, ethylpiperidine, and other amines which have been used to form salts with benzylpenicillins. In the case of compounds (I) which contain a basic nitrogen site in the side chain, acid addition salts may also be formed. Such salts include, for example, inorganic slats such as the sulphate, nitrate, phosphate, borate, thiocyanate, and hydrohalides, e.g. hydrochloride, hydrobromide and hydroiodide, and organic salts such as the acetate, oxalate, tartrate, malate, citrate, succinate, benzoate, ascorbate and methanesulphonate.

Examples of suitable pharmaceutically acceptable esters include those which break down readily in the human body to leave the parent acid, e.g. acyloxyalkyl esters such as acetoxymethyl, pivaloyloxymethyl, α-acetoxyethyl, α-acetoxybenzyl and α-pivaloyloxymethyl esters, and alkoxycarbonylalkyl esters such as methoxy carbonyloxymethyl esters. Other suitable esters of this readily hydrolysable type include lactone, thiolactone, and dithiolactone esters (i.e. compounds of formula (I) wherein the 3-carboxy group is esterified to produce a grouping of formula: ##EQU2## wherein X¹ and Y¹ are oxygen or sulphur and Z¹ is a divalent hydrocarbon group), especially the phthalidyl and substituted phthalidyl esters e.g. 5,6-dimethoxyphthalidyl ester.

The compounds of this invention may be prepared by reacting 6-aminopenicillanic acid or a salt, ester or silyl derivative thereof with an N-acylating derivative of an acid of formula (IV) ##EQU3## in which any reactive substituents may be blocked, wherein R, R¹, R² and R³, are as defined in formula (I) and then, if necessary, carrying out one or more of the following steps (i) removing any silyl groups by hydrolysis or alcoholysis, (ii) converting an ester compound to a free acid or salt (iii) converting a salt to a free acid or a free acid to a salt (iv) removing any blocking groups to release the desired functional substituent (v) converting a free acid compound to an ester compound.

By the term "silyl derivative" used in connection with 6-amino-penicillanic acid (6-APA) we mean the product of the reaction between 6-APA and a silylating agent such as a halotrialkylsilane, halodialkylsilane, a halotrialkoxysilane, a dihalodialkoxysilane or a corresponding aryl or aralkyl silane and compounds such as hexamethyldisilazane. In general, halotrialkylsilanes are preferred, especially trimethylchlorosilane.

A reactive N-acylating derivative of the acid (IV) is employed in the above process. The choice of reactive derivative will of course be influenced by the chemical nature of the substituents in the acid. Thus, when the acid contains only acid stable groups, an acid halide is a suitable N-acylating derivative, preferably the acid chloride.

Such reagents would, however, be avoided when an acid labile group was present in the acid (IV). In such cases a suibable N-acylating derivative is a mixed anhydride. For this purpose particularly convenient mixed anhydrides are the alkoxyformic anhydrides.

However, with both the acid chloride and mixed anhydride N-acylating agents we have found that some racemisation may take place. To minimise such unwanted racemisation, we prefer to use an activated ester as the N-acylating agent. Such activated esters, for example the ester formed with 1-hydroxybenzotriazole or, preferably, N-hydroxysuccinimide, may be prepared in situ by the reaction of the acid with the appropriate hydroxy compound in the presence of a carbodiimide, preferably dicyclohexylcarbodiimide.

Other reactive N-acylating derivatives of the acid (II) include the reactive intermediate formed by reaction in situ with a carbodiimide or carbonyldiimidazole, but the literature on the preparation of semi-synthetic penicillins contains examples of other reactive N-acylating derivatives of acids suitable for coupling to 6-APA.

It will be understood, of course, that where a free acid of type (I) or a salt thereof is desired, it may be convenient to carry out the acylation reaction using an ester of 6-APA, and then to remove the ester group. Vice versa, if an ester is required, it may be convenient to carry out the acylation reaction using 6-APA or a salt thereof and thereafter to esterify the free acid.

In the above process, if it is necessary to block any reactive substituents in the acid (IV), conventional chemical blocking groups are known. Thus, if desired, any free amino groups may be blocked by conversion to benzyloxycarbonylamino groups, or the amino group may be blocked as the nitro group which is later converted to the amino group.

The compounds of this invention may also be prepared by a process which comprises reacting a compound of formula (V) or a salt, ester or silyl derivative thereof. ##SPC2##

wherein R is as defined in formula (I) and in which any reactive substituents may be blocked, with an N-acylating derivative of an acid of formula (VI) ##EQU4## wherein R¹, R² and R³ are as defined in formula (I), and if necessary, carrying out one or more of the following steps (i) removing any silyl groups by hydrolysis or alcoholysis, (ii) converting an ester compound to a free acid or salt thereof (iii) converting a salt to a free acid or a free acid to a salt (iv) removing any blocking groups to release the desired functional substituents (v) converting a free acid compound to an ester compound.

The remarks made earlier with respect to silyl derivatives, N-acylating derivatives, and blocking groups, also apply to this process.

The compounds of this invention wherein R¹ is a group of formula (II) may also be prepared by reacting a compound of formula (VII) or a salt, ester or silyl derivative thereof: ##SPC3##

wherein R, R¹, R³ and R⁴ are as defined in formula (I) and wherein any reactive substituents may be blocked, with cyanate ion, a C₁₋₄ alkyl isocyanate, cyclohexyl isocyanate, a formylating agent or an N-acylating derivative of an acid R⁶ COOH wherein R⁶ is phenyl or an alkyl group having from 1 to 4 carbon atoms, followed, if necessary, by one or more of the following steps (i) removing any silyl groups by alcoholysis or hydrolysis, (ii) converting an ester compound to free acid or salt thereof, (iii) converting a salt to a free acid or a free acid to a salt (iv) removing any blocking groups to release the desired functional substituent, (v) converting a free acid compound to an ester compound.

It will be noted that the above process essentially consists in generating the desired group R² from the free amino group in compound (VII). The reaction of amino compounds with cyanate ion and isocyanates to produce ureas and substituted ureas is well known. Likewise the formylation of amino compounds (e.g. using formic acid and acetic anhydride) is well known. Similarly, the acylation of amino compounds is extremely well known, and suitable N-acylating derivatives of acids have been discussed hereinbefore.

The compounds of this invention are broad spectrum penicillins, i.e. penicillins which not only have activity against Gram-positive bacteria, but also against a number of clinically important Gram-negative organisms. The preferred compounds of this invention are active against such important organisms as Pseudomonas spp. against which the most well known broad-spectrum penicillin (6[(D)α-aminophenylacetamido]penicillanic acid . . . ampicillin) is normally inactive. Moreover the preferred compounds of this invention are about as active as 6[(D)α-carboxy-3-thienylacetamido] penicillanic acid against Pseudomonas spp., this latter compound being the most active of the known penicillins against those organisms. Several of the preferred compounds of this invention have minimum inhibitory concentrations of from 5-12.5 μg/ml against some β-lactamase producing strains of staphylococci, against which the majority of known broad spectrum penicillins are only marginally effective. The preferred compounds of this invention are not greatly serum-bound, and are not markedly inactivated by serum.

The penicillins of this invention show the characteristic lack of toxicity of penicillins generally. They may be administered by parenteral injection. The daily dose will depend on the identity of the penicillin and severity of infectioon. With the preferred compounds of this invention, a suitable average daily dose for an adult would be in the range of 100mg to 5000mg. An average single dose for an adult would be from 20 mg to 500 mg.

The following Examples illustrate the preparation of some of the compounds of this invention: ##SPC4##

In the following Examples amoxycillin is the approved name for 6[D-α-amino-p-hydroxyphenylacetamido]penicillanic acid and ampicillin is the approved name for 6[D-α-aminophenylacetamido] penicillanic acid. Epicillin is the approved name for [D-α-amino -cyclohexa-1,4-dienylacetamido]penicillanic acid. All temperatures are in ° C. All biochromatograms were run in butanol/ethanol/water. All compounds were made by one of the following generally applicable methods.

The majority of the starting materials used in the following Examples are known. However, the following literature references described generally applicable methods which may be used to prepare the starting materials:

    UREIDO-ACIDS                                                                   DAKIN            Amer. Chem. J. 44 54                                          ANDREASCH        Monats. 23. 805                                               NEVILLE, McGEE   Can. J. Chem. 41, 2123-9 (1963)                               WIELAND          Bio. Z. 38, 389, Ann. 3.                                      DAVIS, BLANCHARD J. Amer. Chem. Soc. 51, 1797                                  LEUTHARDT, BRUNNER                                                                              Helv. Chim. Acta. 30, 964-5 (1947)                            SUBSTITUTED UREIDO-ACIDS                                                       BALL, SKINNER, SHIVE                                                                            Texas Rept. Biol. Med. 21(2)                                                   188-75 (1963)                                                 GUANIDINO-ACIDS 1301961/2.2-                                                   KAPFHAMMER, MILLER                                                                              Z. Physiol, Chem. 225, 1-12, (1934)                           RADKA PANT       Ibid 335, 272-4 (1964)                                        FRAMM, KAPELLER  Ann. (1925) 442, 144                                          HABEL            Can. J. Biochem. Physiol. 38, 493 (1960)                      RAMSAY           Ber. 41, 4390                                                 FORMAMIDO-ACIDS                                                                SHEEHAN, YOUNG   J. Amer. Chem. Soc. (1958), 80, 1154                     

Method A

A solution of the guanidino-acid, hydrochloride (5 m. mole) in dry dimethylformamide (5 ml) was added over 10 mins. to a stirred solution of phthalid-3-yl D-α-aminophenyl-acetamidopenicillanate (5 m. mole) and N,N¹ -dicyclohexylcarbodi-imide (5.8 m. mole) at 0° C in dry methylene dichloride.

After stirring at 0° C for 30 mins. and 11/2 hours at ambient temperatures, the mixture was cooled to -10° C and the dicyclohexylurea removed by filtration.

The solution was washed with dilute hydrochloric acid (pH 1.5), water, and brine and the dried solution concentrated to low volume in vacuo to induce crystallisation. The filtered solid was dried under vacuum over phosphorus pentoxide.

Method B

Ureido- (or substituted-ureido-) acid (0.01 mole) in dry acetone (60 ml) at -10° C was treated with triethylamine (ca. 0.015 mole) and iso-butylchloroformate (0.01 mole) and stirred at -10° C for not more than 30 mins. D-α-aminophenylacetamidopenicillanic acid, trihydrate (0.01 mole) in water (60 ml) was treated with triethylamine to give a clear solution (pH 8.4). Acetone (60 ml) was added and the solution cooled to 0° C.

The mixed anhydride solution cooled to -40° C was filtered through Celite into the stirred penicillin solution and the mixture allowed to warm slowly to room temperature over 20 mins.

The acetone was evaporated in vacuo and the aqueous residue washed well with ether and then acidified to pH2 under a layer of ethyl acetate with 5N hydrochloric acid.

The product was obtained either as the free acid by filtration of the aqueous/ethyl acetate mixture or by precipitation from the ethyl acetate solution with potassium or sodium 2-ethylhexoate to give the corresponding alkali-metal salt.

Method Bi) As B, but using N-methylmorpholine instead of triethylamine in the preparation of the mixed anhydride.

Method Bii) As B, but using D-α-amino-(p-hydroxyphenyl)acetamidopenicillanic acid, trihydrate instead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Biii) As Bii) but using N-methylmorpholine instead of triethylamine in the preparation of the mixed anhydride.

Method Biv) As Bi), but using D-α-amino-(3-thienyl)-acetamidopenicillanic acid instead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bv) As Bi), but using D-α-amino-(1,4-cyclohexadienyl)acetamidopenicillanic acid instead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bvi) As Bi), but using ethylchloroformate instead of iso-butyl chloroformate and D-α-aminocyclopropylacetamidopenicillanic acid, trihydrate.

Method Bvii) as Bi), but using D-α-aminovaleramidopenicillanic acid instead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bviii) As Bi), but using ethylchloroformate instead of isobutyl chloroformate, and D-α-amino-(2-thienyl)-acetamidopenicillanic acid instead of D-α-aminophenylacetamidopenicillanic acid, trihydrate.

Method Bix) As Bi), but using D-α-amino-β-phenylpropionamidopenicillanic acid instead of D-α-aminophenylacetamidopenicillanic acid. (No Method C)

Method D

Amino-penicillin (5 m. moles) in dry dimethylformamide (100ml) was treated with triethylamine (12 m. moles) and stirred to give a clear solution. Sulphur trioxide-triethylamine complex (6 m. moles) was added portionwise over 5 mins. at room temperature and stirred for 1 hour. A solution of potassium 2-ethyl hexoate (ca 15 m. moles) in dry acetone (150ml) was added and a white solid separated.

After further dilution with acetone (200ml) the solid was filtered, washed with acetone and then stirred in dry ether for 20 mins. to remove any residual dimethylformamide. The solid was filtered and dried in vacuo.

Method E

Anhydrous D-α-aminophenylacetamidopenicillanic acid (5 m. mole) in dry methylene dichloride (50 ml) was treated with triethylamine (˜10 m. mole) to give a clear solution. Trimethylsilyl chloride (10 m. mole) was added and the mixture refluxed under nitrogen for 1 hour, then cooled to 0° C.

α-Guanidino-acid (5 m. mole) was dissolved in dry dimethylformamide 55ml) and dry dimethylformamide (5ml.) and dry methylene dichloride (50 ml) added, cooled to 0° C and stirred for 5 mins. with dicyclohexyl-carbodi-imide (5.5 m. mole). The bis-trimethylsilylated penicillin was added and stirred at 0° C for 1 hour. The mixture was then cooled to -20° C and the dicyclohexylurea removed by filtration. The filtrate was evaporated to dryness in vacuo and the residue dissolved in acetone (20ml)/water (20 ml) and the pH adjusted to 2.5 with 5N hydrochloric acid. After stirring at pH 2.5 for 25 mins. The acetone was removed in vacuo and any solid filtered off. The residual aqueous solution was freeze dried and the resultant solid treated with water at pH 2. The product was filtered and dried.

Method F

Dicyclohexylcarbodi-imide (5.5 m. mole) was added to a stirred solution of N-substituted-amino acid (5 m. mole) in dry acetone (20 ml) at 0° C. The mixture was stirred for 15 mins. at 0°-5° C and then left in the refrigerator overnight.

D-α-Aminophenylacetamidopenicillanic acid, trihydrate (5 m. mole) was dissolved in acetone (10 ml)/water (10 ml) with triethylamine (0.7 ml) and the hydroxysuccinimide ester filtered in, through Celite. After stirring for 45 mins. the acetone was removed in vacuo, leaving a gelatinuous mass. Acidification with 5N hydrochloric acid in aqueous ethyl acetate gave the product as the free acid (sometimes only after concentration of the ethyl acetate layer and treatment with ether) or as the salt by treatment of the washed and dried ethyl acetate layer with sodium or potassium 2-ethyl hexoate.

Method Fi) As F, but the hydroxysuccinimide ester formed in dry dimethylformamide (or dimethylformamide diluted with acetone).

Method Fii) As F, but the hydroxysuccinimide ester formed in dry 1,2-dimethoxyethane.

Method Fiii) As Fi), but the penicillin dissolved in acetone/chloroform, the product coming out of solution as the amine salt.

EXAMPLE 1 D-α-[D-β-(p-Hydroxyphenyl)-α-ureidopropionamido]phenylacetamido penicillanic acid

(R = Ph; R¹ = -p--HO--PhCH₂ ; R³ = H; R² = NHCONH₂ ; M = H; α¹ = D).

Prepared by method Bi), from D-β-(p-Hydroxyphenyl)-α-ureidopropionic acid.

Yield: 68%. νmax (KBr): 3550, 1770, 1650, 1515, 1230 and 700cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.44 (3H. s. gem methyl); 1.57 (3H s. gem methyl); ˜2.8 (2H. m. -CH₂ CH<); 4.29 (1H. s. C-3 proton); ˜4.5 (1H. m. -CH₂ CH<); 5.35 - 5.87 (5H. m. β-lactams, Ph CH--; NHCONH₂ *) 6.27 (1H. d. NHCONH₂ *); 6.67 ##SPC5##

7.30 (5h. broad s. pH CH<); 8.47 (1H. m. -CONH*); 9.12 (1H. m. -CONH*).

NH₂ OH Assay: 101%. Biochromatography: 1 zone at R_(f) 0.32.

EXAMPLE 2 D-α-[D-β-(-p-Hydroxyphenyl)-α-ureidopropionamido]-(-p-hydroxyphenyl) acetamidopenicillanic acid.

(R = --p--HO--Ph; R¹ = --p--HO--PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ = D).

Prepared by method Bi), from D-β-(-p-hydroxyphenyl)β-ureidopropionic acid and amoxycillin.

Yield: 74%. νmax (KBr): 3350 (broad), 1770, 1650, 1515, 1230 and 840 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H. s. gem methyl); 1.57 (3H. s. gem methyl); ˜2.8 (2H. m. --CH₂ CH<); 4.3 (1H. s. C-3 proton); ˜4.4 (1H. m. CH₂ CH<); 5.3 - 5.85 (5H. m. β-lactams, Ph CH-; NHCONH₂ *); 6.25 - 7.30 (8H. m. aromatics protons); 8.47 (1H. m. CONH*-); 9.12 (1H. m. -CONH*-).

NH₂ OH assay: 94%. Biochromatography: 1 zone at R_(f) = 0.18.

EXAMPLE 3 D-α-[D-β-(p-Hydroxyphenyl)-α-ureidopropionamido](3-thienyl)actamido penicillanic acid. ##SPC6##

Prepared by method Bii), from D-β-(p-hydroxyphenyl)-α-ureidopropionic acid and α-amino-(3-thienyl) acetamido pencillanic acid.

Yield: 72% νmax (KBr): 3350 (broad); 1770, 1650, 1515, 1230, 845 and 780cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.45 (3H. s. gem methyl); 1.58 (3H. s. gem methyl); ˜2.8 (2H. m. -CH₂ CH<); 4.3 (1H. s. C-3 proton), ˜4.4 (1H. m. -CH₂ CH<); 5.35 - 5.90 (5H. m. β-lactams, Ph -CH-; NHCONH₂ *); 6.25 (1H. m. NH CONH₂); 6.5 - 7.6 (7H. m. aromatics); 8.5 (1H. d. CONH*); 9.1 (1H. d.CONH*).

NH₂ OH assay: = 96% Biochromatography: 1 zone at R_(f) = 0.31

EXAMPLE 4 D-α-[DL-β-(-p-Nitrophenyl)-α-ureidopropionamido]-(-p-hydroxyphenyl)acetamidopenicillanic acid.

(R = p--HO--Ph; R¹ = p--NO₂ --PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ = DL).

Prepared by method Bii), from DL-β-(-p-Nitrophenyl)-α-ureidopropionic acid and amoxycillin.

Yield: 60%. νmax (KBr): 3350, 1770, 1650, 1514 and 1230 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H. s. gem methyl); 1.55 (3H. s. gem methyl); 3.0 (2H. m. CH₂ CH<); 4.05 (1H. s. C-3 proton); ˜4.60 (1H. m. -CH₂ CH<); 5.25 - 5.80 ##SPC7##

6.2 (1h. m. NHCONH₂ *); 6.5 - 8.2 (8H. m. aromatics); 8.50, 9.00 (2 × 1H.m.CONH*).

NH₂ OH assay: 86.5% Biochromatography: 1 zone at R_(f) = 0.44

EXAMPLE 5 Sodium D-α-[DL-γ-Methylthio-α-ureidobutyramido]phenylacetamidopenicillanate.

(R = Ph; R¹ = CH₃ S (CH₂)₂ -; R³ =H; R² = NHCONH₂ ; M=Na; α¹ = DL).

Prepared by Method B, from N-Carbamoyl-DL-methionine and ampicillin, isolated as the sodium salt after treatment with sodium 2-ethylhexoate.

Yield: 51%. νmax (KBr): 3320 (broad), 1775, 1650, 1530, 1310, 1230 and 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.48 (3H. s. gem methyl); 1.58 (3H. s. gem methyl); 1.4 - 2.2 (2H. m. -SCH₂ CH₂ CH<); 2.05 (3H. d. MeS-); 2.3 - 2.7 (2H. m. SCH₂ --); 4.24 (1H. s. C-3 proton); 4.1 - 4.6 (1H. m. SCH₂ CH₂ CH<); 5.2 - 5.9 (5H. m. β-lactams. PhCH< and --COHN₂ *); 6.4 (1H. m. CONH*-); 7.40 (5H. m. aromatics); 8.60 and 9.00 (2 × 1H. d. CONH*-).

NH₂ OH assay: 93.8% Biochromatography: 1 zone at R_(f) = 0.36.

EXAMPLE 6 Sodium D-α-[DL-γ-Methylthio-α-ureidobutyramido](p-hydroxyphenyl)acetamido penicillanate.

(R = p-HO-Ph; R¹ = CH₃ S (CH₂)₂ ; R³ = H; R² = NHCONH₂ ; M=Na; α¹ =DL).

Prepared by method Bii), from M-Carbamoyl-DL-methanionine and amoxycillin, isolated as the sodium salt after treatment with sodium 2-ethylhexoate.

Yield: 43%. νmax (KBr): 3350 (broad), 1770, 1650, 1510, 1235cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.45 (3H. s. gem methyl); 1.57 (3H. s. gem methyl); 1.4 - 2.2 (2H. m. -SCH₂ CH₂ CH<); 2.05 (3 H. d. MeS-); 2.3 - 2.7 (2H. m. SCH₂ -); 4.24 (1H. s. C-3 proton); 4.1 - 4.6 (1H. m. SCH₂ CH₂ CH<); 5.2 - 5.9 (5H.m.β-lactams, PH CH< and CONH₂ *); 6.4 (1H. m. CONH*-); 6.3 - 7.34 (4H. m. aromatics); 8.60 and 8.90 (2 × 1H.d.CONH*--).

NH₂ OH assay: 85.5% Biochromatography: 1 zone at R_(f) = 0.29.

EXAMPLE 7 Sodium D-α-[DL-α-formamido-γ-methylthiobutyramido]phenylacetamido pencillanate

(R = Ph; R¹ = CH₃ S (CH₂)₂ ; R³ = H; R² = NHCHO; M=Na; α¹ = DL).

Prepared by Method B, using N-Formyl-DL-methionine and ampicillin, isolated as the sodium salt after treatment with sodium 2-ethyl hexoate.

Yield: 63%. νmax (KBr): 3300 (broad), 1780, 1732, 1645, 1525, 1302, 1225 and 700 cm.sup.⁻¹ δ[(CD₃)₂ SO]: 1.46 (3H. s. gem methyl); 1.55 (3H. s. gem methyl); 1.7 - 2.2 (2H. m. CH₃ SCH₂ CH₂ CH<); 2.05 (3H. d. CH₃ S-); 2.3 - 2.7 (2H. m. CH₃ S CH₂ CH₂); 4.23 (1H. s. C-3 proton); 4.70 (1H. m. -CH¹ NHCHO); 5.3 - 5.9 (3H. m. β-lactams and PhCH<); 7.37 (5H. m. aromatics); 8.08 (1H. s. NHCHO); 8.31, 8.60 and 9.00 (3 × 1H. d. -COHN-*).

NH₂ OH assay: 100% Biochromatogram: Single zone at R_(f) = 0.40.

EXAMPLE 8 D-α-[DL-β-(p-Chlorophenyl)-α-ureidopropionamido]-(p-hydroxyphenyl)acetamidopenicillanic acid.

(R = p-HO-Ph; R¹ =p-Cl-PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=Na; α¹ = DL).

Prepared by Method Bii) from DL-β-(P-Chlorophenyl-α-ureidopropionic acid and amoxycillin.

Yield: 45%. νmax (KBr): 3360, 1770, 1650, 1514 and 1230 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H. s. gem dimethyl); 1.58 (3H. s. gem dimethyl); 2.86 (2H. m. -CH₂ CH<); 4.26 (1H. s. C-3 proton); 4.55 (1H. m. -CH₂ CH<); ##SPC8##

6.24 (1h. m. NHCONH₂ *); 6.78 and ##SPC9##

8.53 (1H. m. CONH); 9.04 (1H. m. CONH).

Biochromatagraphy: R_(f) = 0.42 Analysis: C₂₆ H₂₈ N₅ O₇ SCl required: C, 52.93; H, 4.75; N, 11.87; S, 5.43; Cl, 6.02. Found: C, (50.23); H, 4.79; N, 11.13; S, 5.32; Cl, 5.97.

EXAMPLE 9 D-α-[DL-β-(p-Fluorophenyl)-α-ureidopropionamido]phenylacetamido penicillanic acid

(R = Ph; R¹ = p-F-PhCH₂ ; R³ = H; R² = NHCONH₂ ; M = H; α¹ = DL)

Prepared by method B from DL-β-(p-fluorophenyl)-α-ureidopropionic acid and ampicillin.

Yield: 42%. νmax (KBr): 3360, 1773, 1651, 1510, 1226 and 720 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H. s. gem methyl); 1.57 (3H. s. gem methyl); 1.57 (3H. s. gem methyl); 2.87 (2H. m. CH₂ CH<); 4.24 (1H. s. C-3 proton); 4.56 (1H. m. CH₂ CH<); 5.65 (5H. m. β-lactams, α-proton, -NHCONH₂ *), 6.27 (1H. d. NHCONH₂ *); 7.24 (9H. m. Ph CH<); ##SPC10##

9.17 (1h. m. CONH*).

Bichromatography: 1 Spot at R_(f) = 0.42.

EXAMPLE 10 D-α-[DL-β-(p-Chlorophenyl)-α-ureidopropionamido]phenylacetamido penicillanic acid

(R = Ph; R¹ = p-Cl-PhCH₂ ; R³ = H: R² = NHCONH₂ ; M=N; α¹ = DL).

Prepared by Method B, from DL-β-(p-chlorophenyl)-α-ureidopropionic acid.

Yield: 38% νmax (KBr): 3360, 1770, 1650, 1514 and 1230 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.40 (3H. s. ge m methyl); 1.56 (3H. s. gem methyl); 2.85 (2H. m. --CH₂ CH<); 4.25 (1H. s. C-3 proton); 4.55 (1H. m. -CH₂ CH<); ##SPC11##

6.25 (1h. m. NCONH₂); 7.30 (9H. m. aromatic protons); 8.53 (1H. m. CONH); 9.00 (1H. m. CONH). NH₂ OH Assay: 94% Biochromatography: 1 zone at R_(f) = 0.59.

EXAMPLE 11 Sodium D-α-[DL-β-(p-Nitrophenyl)-α-ureidopropionamido]phenylacetamidopenicillanate.

(R = Ph; R¹ = p-NO₂ -PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=Na; α¹ = DL).

Prepared by method B, from DL-β-(p-Nitrophenyl)-α-ureidopropionic acid and ampicillin; isolated as the sodium salt by treatment with sodium 2-ethylhexoate.

Yield: 55%. νmax (KBr); 3350, 1770, 1650, 1514 and 1230 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H. s. gem methyl); 1.60 (3H. s. gem methyl); 3.0 (2H. m. -CH₂ CH<); 4.20 (1H. s. C-3 proton); 4.7 (1H. m. -CH₂ CH<); ##SPC12##

6.2 (1h. m. NHCONH₂ *); 7.18 - 8.25 (9H. m. aromatics); 8.50, 9.05 (2 × 1H. m. CONH*).

NH₂ OH assay: 93.9% Bichromatography: 1 zone at R_(f) = 0.5.

EXAMPLE 12 D-α-(D-p-Phenyl-α-ureidopropionamido)-(p-hydroxyphenyl)acetamidopenicillanic acid

(R = p-HO-Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCONH₂ ; M=H; α¹ = D).

Prepared by method Biii), using D-β-phenyl-α-ureidopropionic acid and amoxycillin.

Yield: 63% M.P. 235°-238°C. νmax (KBr): 3360 (broad), 1740, 1650, 1520 and 1230 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.47 (3H. s. gem methyl); 1.60 (3H. s. gem methyl); 2.95 (2H. m. Ph.CH₂.CH<); 4.27 (1H. s. C-3 proton); 4.60 (1H. m. PhCH₂ CH<); ##SPC13##

7.25 (5h. s. benzyl aromatics), 8.50 (1H. d. -CONH-*); 9.00 (1H. d. -CONH-*).

Hydroxylamine Assay: 107.7%. Biochromatography: One zone at R_(f) = 0.35. Analysis for C₂₆ H₂₉ N₅ O₇ S; Required (%) C, 56.22; H, 5.23; N, 12.61; S, 5.77. Found (%) C, (55.33); H, 5.44; N, 11.99; S, 5.44.

EXAMPLE 13 D-α-(DL-α-acetamido-β-phenylpropionamido)-phenyl-acetamido-penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCOCH₃ ; M = H; α¹ = DL)

Prepared by method Bi, from DL-α-acetamido-β-phenylpropionic acid and ampicillin.

Yield: 94%. νmax (KBr): 33.60 (broad) 1774, 1648, 1511, 1215 and 701cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.43 (3H. s. gem methyl); 1.56 (3H. s. gem methyl); 1.77 (3H. s. NHCOCH₃); 2.7-3.2 (2H.m. PhCH₂ CH<) 4.24 (1H.s. C-3 proton); 4.6-4.9 (1H.m. PhCH₂ CH<); 5.62 (3H.m. β-lactams, PhCH<); 7.38 (10H.m. PhCH<; Ph CH₂ OH<); 8.0-9.3 (3H.m. removable in D₂ O, -COHN-) Hydroxyalamine Assay: 62.1%. Biochromatography: 1 zone at R_(f) = 0.26. Analysis: C₂₇ H₃₀ N₄ O₆ S required: C, 60.22; H, 5.58; N, 10.41; S, 5.95. Found: C, (57.45); H, 5.69; N, 9.98; S, 6.03.

EXAMPLE 14 D-α-[DL-α-(3-Methylureido)-β-phenyl-propionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = NHCONHCH₃ ; M = H; α¹ = DL)

Prepared by method Bi, starting from DL-α-(3-methylureido)-β-phenylpropionic acid ampicillin.

Yield: 75%. νmax (KBr): 1775, 1637, 1560, 1490, 1297, 1219 and 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl); 1.57 (3H.s. gem methyl); 2.52 (3H.s. NHCONHCH₃); 2.90 (2H.m. PhCH₂ CH<); 4.27 (1H.s. C-3 proton); 4.63 (1H.m. PhCH₂ CH<); 5.64 (3H.m. β-lactams, PhCH<); 7.32 (10H.m. PhCH₂ CH<); PhCH<); 8.63 (1H.m. removable in D₂ O,--CONH--); 9.18 (1H.m. removable in D₂ O,--CONH--); ca 6.25 (broad signal due to --NH CONH--).

Biochromatography: 1 zone at R_(f) = 0.47 Analysis: C₂₇ H₃₁ N₅ O₆ S required: C, 58.48, H, 5.60; N, 12.64; S, 5.78 Found: C, (56.78); H, 5.59; N, 12.73; S, 5.05.

EXAMPLE 15 D-α-[DL-β-(-p-Fluorophenyl)-α-ureido-propionamido]-(-p-hydroxyphenyl)acetamido-penicillanic acid

(R = -p-HO-Ph; R¹ = -p-F-PhCH₂ ; R³ = H; R² = NHCONH₂ ; M = H, α¹ = DL)

prepared by method Bii, from DL-β-(-p-fluorophenyl)α-ureidopropionic acid and amoxycillin.

Yield: 48% νmax (KBr): 3360, 1764, 1650, 1510, 1224, and 838cm.sup.⁻¹ δ[(CD₃)₂ SO]: 1.43 (3H.s. gem methyl); 1.54 (3H.s. gem methyl); 2.88 (2H.m. -CH₂ CH<); 4.14 (1H.s. C-3 proton); 4.2-4.8 (1H.m. CH₂ CH<); 5.3-7.5 ##SPC14##

δ= 8.3-9.2 (2h.m. -CONH₂ -*)

Hydroxylamine Assay: 84.7%. Biochromatography 1 zone at R_(f) = 0.41.

EXAMPLE 16 D-α-[D-β-Phenyl-α-(n-valeramido)-propionamido]phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCO (CH₂)₃ CH₃ ; M = H; α¹ = D)

Prepared by method Bi, from N-valeroyl-D-β-phenylalanine

Yield: 19% νmax (KBr): 3290 (br), 1773, 1635, 1525, 1300, 1224, 733, 702cm.sup.⁻¹. δ[(CH₃)SO]; 0.82 (3H.m. (CH₂)₃ CH₃), 1.0-1.7 (4H.m. CH₂ CH₂ CH₂ CH₃) 1.45 (3H.s. gem methyl); 1.58 (3H.s. gem methyl); 2.09 (2H.m. CH₂ (CH₂)₂ CH₃), 3.00 (2H.m. PhCH₂), 4.28 (1H.s. C-3 proton), 4.78 (1H.m. PhCH₂ CH<) 5.4-5.9 (3H.m. β-lactams and PhCH<), 7.2-7.5 (10H.m. aromatic protons), 8.05 (1H.d. --CONH--), 8.47 (1H.m. CONH), 9.13 (1H.m. --CONH--). NH₂ OH Assay: 90%. Biochromatography: Single zone R_(f) = 0.70. Analysis: Found: C, 60.96; H, 6.05; N, 9.46; S, 5.65% C₃₀ H₃₆ N₄ O₆ S requires C, 62.10; H, 6.21; N, 9.66; S, 5.52.

EXAMPLE 17 D-α-[D-β-Phenyl-α-pivaloylaminopropionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ : R³ = H; R² = -NHCOC(CH₃)₃ ; M = H; α¹ = D)

Prepared by method Bi, from α-t-Butyramido-D-β-phenyl propionic acid.

Yield: 6%. νmax (KBr); 3350(br), 1772, 1639, 1517, 1300, 1212, 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 0.97 (9H.s. (CH₃)₃), 1.39 (3H.s. gem methyl), 1.52 (3H.s. gem methyl), 2.97 (2H.m. PhCH₂), 4.20 (1H.s. C-3 proton), 5.35-5.85 (4H.m. β-lactams and PhCH<), 4.65 (1H.m. PhCH₂ CH<), 7.25 (5H.s. phenyl protons), 7.39 (5H.m. phenyl protons), 7.2-7.6 (1H.m. --NHCO--*), 8.50 (1H.d. --NHCO--*), 9.27 (1H.d. --NH--CO--).

NH₂ OH Assay: 102%. Biochromatography: 0.71.

EXAMPLE 18 D-α-[D,L-α-Benzamido-β-phenylpropionamido] -phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCO Ph; M = H; α¹ = D,L)

Prepared by method Bi, from α-Benzamido-D,L-β-phenylpropionic acid.

Yield: 22%. νmax (KRr): 3300, 1775, 1635, 1522, 1302, 122, 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H.s. gem methyl), 1.57 (3H.s. gem methyl) 3.11 (2H.m. PhCH₂), 4.24 (1H.s. C-3 proton), 5.00 (1H.m. PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<), 7.2-8.0 (10H.m. aromatic protons), 8.65, 8.80 and 9.20 (3 × 1H.d. -NHCO-*).

NH₂ OH Assay: 88%. Biochromatography: Single zone 0.70.

EXAMPLE 19 D-α-[D,L-γ-Phenyl-α-ureidobutyramido] -p-hydroxyphenylacetamido penicillanic acid

(R = p HO-Ph; R¹ = PhCH₂ CH₂ -; R³ = H; R² = -NHCONH₂ M = H; α¹ = D,L)

Prepared by method Biii, from α-ureido-D,L-α-phenylbutyric acid.

Yield: 35%. νmax (KBr): 3315(br), 1770, 1650, 1510, 1454, 1227, 842, 703cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.42 (3H.s. gem methyl), 1.52 (3H.s. gem methyl), 2.00 (2H.m. PhCH₂ CH₂), 2.52 (2H.m. PhCH₂ CH₂), 4.1-4.4 (1H.m. PhCH₂ CH₂ CH<), 4.25 (1H.s. C-3 proton), 5.3-5.8 (3H.m. β-lactams and PhCH<), 7.25 (9H.m. aromatic protons, 6.34, 6.73 (2 × 1H.d. NHCO*), 8.32-9.10 (3H.m. --CONH--* and --CONH*₂)

EXAMPLE 20 D-α-[D,L-α-Formamido-β-phenylpropionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = -NHCHO; M = H; α¹ = D,L)

Prepared by method Bi from N-Formyl-D,L-phenylalanine.

YIELD: 55%. νmax (KBr): 3242(br), 1771, 1638, 1522, 1379, 1300, 1226, 731, 701cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.46 (3H.s. gem methyl), 1.59 (3H.s. gem methyl); 2.88 (2H.m. PhCH₂ CH<), 4.21 (1H.s. C-3 proton), 4.83 (1H.m. PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<), 7.2-7.6 (10H.m. aromatic protons), 7.97 (1H.s. CHO), 2.27, 2.70 and 9.11 (3 × 1H.d. --NHCO*--).

NH₂ OH Assay: 79%. Biochromatography: Single zone R_(f) 0.52.

EXAMPLE 21 D-α-[D-β-Phenyl-α-propionamido -propionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = NHCOCH₂ CH₃ ; M = H; α¹ = D)

Prepared by method Bi from α-Propionamido-D-β-phenylpropionic acid.

Yield: 13%. νmax (KBr); 3229(br), 1770, 1637, 1524, 1226, 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 0.89 (3H.t. COCH₂ CH₃), 1.42 (3H.s. gem methyl), 1.60 (3H.s. gem methyl), 1.98 (2H.m. -NHCH₂ CH₃), 2.90 (2H.m. PhCH₂ CH<), 4.21 (1H.s. C-3 proton), 4.70 (1H.m. PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<), 7.2-7.6 (10H.m. aromatic protons), 8.01, 8.42 and 9.10 (3 × 1H.d. --CONH*--).

NH₂ OH Assay: 75%. Biochromatography: Single zone R_(f) 0.58.

EXAMPLE 22 D-α-[D-α-Isobutyramido-β-phenylpropionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ -; R³ = H; R² = NHCOCH(CH₃)₂ ; M = H; α¹ = D).

Prepared by method Bi, from α-isobutyramido-D-β-phenyl propionic acid

Yield: 18% νmax (KBr): 3300(br), 1771, 1638, 1526, 1300, 1222, 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 0.85 (6H.t. CH(CH₃)₂, 1.41 (3H.s. gem methyl), 1.56 (3H.s. gem methyl), 2.97 (2H.m. PhCH₂ CH<), 4.25 (1H.s. C-3 proton), 4.70 (2H.m. CH(CH₃)₂ and PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<), 7.1-7.6 (10H.m. aromatic protons), 7.97, 8.47 and 9.13 (3 × 1H.d. -- NHCO--*).

NH₂ OH Assay: 103% Biochromatography: Single zone R_(f) 0.66

EXAMPLE 23 D-α-[D-γ-Methylthio-α-ureidobutyramido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = CH₃ S(CH₂)₂ -; R³ = H; R² = -NHCONH₂ ; M = H; α¹ = D)

Prepared by method Bi, from N-Carbamoyl-D-methionine.

Yield: 43%. νmax (KBr): 3320(br), 1775, 1650, 1530, 1310, 1230, and 702cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.48 (3H.s. gem methyl), 1.61 (3H.s. gem methyl), 1.4-2.2 (2H.m. -SCH₂ CH₂ CH<), 2.1 (3H.s.CH₃ S--), 2.3- 2.7 (2H.m. --SCH₂ CH₂ CH) 5.2-5.9 (5H.m.β-lactams, PhCH and --CONH₂ *) 6.4 (1H.m. --CONH--*)

NH₂ OH Assay: 98%. Biochromatography: Single zone R_(f) 0.34. Analysis: Found: C, 49.33; H, 5.64; N, 12.94; S, 11.59% C₂₂ H₂₉ N₅ O₆ S₂ requires C, 50.48; H, 5.54; N, 13.38; S, 12.24%.

EXAMPLE 24 D-α-[D,L-α-Methyl-α-ureidovaleramido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = (CH₃)₂ CHCH₂ -; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = D,L)

Prepared by method B from α-methyl-D,L-α-ureido valeric acid.

Yield: 36% νmax (KBr) 3325(br), 1775, 1723, 1650, 1530, 1310, 1222, 702cm.sup.⁻¹ δ[(CD₃)₂ SO]: 0.84 and 0.95 (2 × 3H.s. CH(CH₃)₂), 1.45 (3H.s. gem methyl), 1.61 (3H.s. gem methyl), 0.78-2.0 (3H.m. CH₂ CH(CH₃)₂), 4.25 (1H.s. C-3 proton), 4.0-4.6 (1H.m. --CHNHCONH₂), 5.3-5.9 (5H.m. β-lactams, PhCH< and CONH₂ *), 6.2 (1H.d. --CONH--), 7.2-7.6 (5H.m. aromatic protons), 8.48 and 9.04 (2 × 1H.d. --CO--NH--* )

NH₂ OH Assay: 79%. Biochromatography: Single zone R_(f) 0.44.

EXAMPLE 25 D-α-[D,L-α-Formamido-β-(p-hydroxyphenyl) propionamido]phenylacetamido penicillanic acid.

(R=Ph; R'=pHO-PhCH₂ -; R³ = H; R² =NHCHO; M=H; α'=D,L)

Prepared by method B from N-Formyl-D,L-Tyrosine

Yield: 23%. νmax (KBr): 3290 (br.), 1773, 1735, 1650, 1518, 1378, 1230 and 701 cm.sup.⁻¹. δ[(CD₃)₂ SO] 1.45 (3H.s gem methyl), 1.62 (3H.s. gem methyl), 2.7- 3.2 (2H.m. PhCH₂ --), 4.24 (1H.s. C3 proton), 4.8 (1H.m. -CHNHCHO), 5.35-5.96 (3H.m. βLactams and Ph CH), 6.7 and 7.0 (2 × 2H.m. pHO--C₆ H₄ --), 7.36 (5H.m. Aromatic protons), 7.92 (1H.s--NHCHO), 8.2 (1H.d. -- CONH--*) 8.7 and 9.12 (2 × 1H.m.--CONH--*).

NH₂ OH Assay: 105%. Biochromatography: Single Zone R_(f) 0.39.

EXAMPLE 26 Dα-[D-α-Formamido -γ -methylthiobutyramido] -phenylacetamido penicillanic acid.

(R=Ph; R'=CH₃ S (CH₂)₂ -; R³ =H; R² =-NHCHO; M=H; α'=D)

Prepared by method Fii from N -Formyl-D-methionine.

Yield: 5.8% νmax (KBr): 3300 (br), 1780, 1732, 1645, 1525, 1302, 1225, 700 cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.46 (3H.s. gem methyl), 1.55 (3H.s. gem methyl), 1.7-22 (2H.m. CH₃ SCH₂ CH₂ CH<), 2.0 (s. 15-17% of L-CH₃ SCH₂ CH₂ CH<) 2.1 (S. 83-5% of D-CH₃ SCH₂ CH₂ CH<), 2.3-2.7 (2H.m. CH₃ SCH₂ CH₂ CH₂), 4.23 (1H.s. C₃ proton), 4.70 (1H.m. -CHNHCHO), 5.3-5.9 (3H.m. βlactams, and PhCH<), 7.37 (5H.m. aromatic protons), 8.08 (1H.s. -NH-CHO), 8.31, 8.59 and 9.02 (3 × 1H.s. -CONH-

NH₂ OH Assay: 102%. Biochromatogram: Single zone R_(f) 0.41.

EXAMPLE 27 D-α -[D-γ-Methyl-α-ureidovaleramide]-phenylacetamido penicillanic acid.

(R=PH; R'= (CH₃)₂ CHCH₂ ; R³ =H; R² =NHCONH₂ ; M=H; α'=D) Prepared by method Fi) from Y-methyl-D 1α-ureido acid.

Yield: 28% M.p: 168°-170°C (Decomp.). νmax (KBr): 3315 (br.), 1775, 1730, 1650, 1530, 1310, 1220, 700 cm⁻ ¹. δ[(CD₃)₂ SO]: 0.85 and 0.92 (6H.d. CH(CH₃)₂), 1.50 (3H.s. gem methyl) 1.66 (3H.s. gem methyl), 0.8-2.0 (3H.m. CH₂ CH (CH₃)₂), 4.28 (1H.s. C3 proton), 4.1-4.5 (1H.m. CH NHCONH₂), 5.3-5.9 (5H.m.β lactams PhCH<and CONH₂ *), 6.22 (1H.d.--CONH*--), 7.36 (5H.m. aromatic protons), 9.07 and 9.40 (2 × 1H.d.-- CO NH*--)

NH₂ OH Assay: 95%. Biochromatogram: Single zone R_(f) 0.45 Analysis: Found: C, 54.71; H, 6.15; N, 13.78; S, 6,49%. C₂₃ H₃₁ N₅ O₆ S requires C, 54.65, H, 6.14; N, 13.86; S, 6.34%.

EXAMPLE 28 D-α[L-γ-Methylthio-α-ureidobutyramido] phenylacetamido penicillanic acid (R=Ph; R'= (CH₃) S (CH₂)₂ --; R² =H; R² = NHCONH₂ ; M=H, α'=L).

Prepared by method Fi) from N-Carbamoyl-L-methionine.

Yield: 19% M.p.: 163°-6°C (Decomp.). νmax (KBr): 3350 (br), 1775, 1650, 1522, 1305, 1220, 702 cm⁻ ¹. δ[(CD₃)₂ SO]: 1.46 (3H.s gem methyl), 1.60 (3H.s. gem methyl) 1.5-2.2 (₂ H.m. CH₃ CH₂ CH₂ CH` ), 2.00 (3H.s. CH₃ S) 2.2-2.6 (2H.m. CH₃ SCH₂ CH₂ CH), 4.27 (1H.s. C3 proton), 1.38 (1H.m. CH₃ CH₂ CH₂ CH<), 5.60 (5H.m.

β lactams, PhCH<, -- CONH*₂), 7.39 (5H.m. aromatic protons), 6.34 (1H.m. -- CHCONH*₂), 9.12 and 9.48 (2 × 1H.d -- CONH*--)

NH₂ OH Assay: 100% Biochromatogram: Single zone R_(f) 0.38 Analysis: Found: C, 49.64; H, 5.98; N, 13.08; S, 11.72%. C₂₂ H₂₉ N₅ O₆ S₂ requires C 50.48; H, 5.54; N, 13.38; S, 12.24%.

EXAMPLE 29 Triethylammonium D-α-[D-β-(3-Indolyl)-α-ureido- propionamido] phenylacetamido penicillanate. ##SPC15##

from N-Carbamonyl-D-Tryptophan Carbamonyl-D-Tryptophan

Yield: 52% M.p.: 200-3°C (Decomp). νmax (KBr): 3350 (br), 1767, 1660, 1640, 1610, 1530, 1458, 1392, 749 cm⁻ ¹. δ[(CD₃)₂ SO]: 1.11 (3H.t. OCH₂ CH₃), 1.42 (3H.s. gem methyl), 1.53 (3H.s. gem methyl), 2.7-3.2 (4H.m.--CH₂ CH<and OCH₂ CH₃), 4.00 (1H.s C₃ proton), 4.59 (1H.m. CH₂ CH<) 5.3-5.9 (5H.m. β lactams, PhCH< and CONH*₂), 6.9-7.7 (10H.m. aromatic protons), 6.29, 8.53 and 8.97 (3 × 1H.d. --NHCO*), 10.84 (1H.s. indolyl NH*).

NH₂ OH Assay: 94% Biochromatogram: single zone R_(f) 0.30 Analysis: Found: C, 59.38; H, 6.65; N, 14.35; S, 4.60% C₃₄ H₄₅ N₇ O₆ S requires C, 60.07; H, 6.67; N, 14.42, S, 4.72%.

EXAMPLE 30 Triethylammonium D-α-[D,L-α-formamido-β-(3-indoly)-propionamido] phenylacetamido penicillinate, ##SPC16##

from N-Formyl-D,L-tryptophan, the product crystallising on dilution with ether.

Yield: 50%. νmax (KBr): 3310 (br), 1772, 1770, 1665, 1530, 1458, 1388, 1218, 748 cm⁻ ¹. δ [ (CD₃) SO]: 1.17 (3H.t.OCH₂ CH₃), 1.44 (3H.s gem methyl), 1.58 (3H.s. gem methyl), 2.8-3.3 (4H.m. OCH₂ CH₃ and CH₂ CH<), 4.12 (1H.s. C₃ proton), 4.90 (1H.m. CH₂ CH<), 5.3-5.9 (3H.m. β-lactams and Ph CH<), 7.0-7.8 (11H.m. 10 aromatic protons and NHCONH₂) 8.00 (1H.S. CHO), 8.0-9.0 (4H.m. 2 ×--NHCO* and --CONH₂ *), 10.80 (1H.s indolyl NH*)

NH₂ OH Assay: 183%. Biochromatogram: Single zone R_(f) 0.36.

EXAMPLE 31: Triethylammonium -α-[D-Y-carbamoyl-α-Ureido butyrylamido] phenylacetamido penicillanate.

(R = Ph; R' = H₂ NCO (CH₂)₂ -; R³ =H; R² = -NHCONH₂ ; M=HN (CH₂ CH₃)₃ ; α'=D.

Prepared by method Fi) from N-Carbamoyl-D-glutamine, the product crystallising on dilution with ether.

Yield: 77%. νmax (KBr): 3400 (br.), 1773, 1698, 1660, 1603, 1532, 1458, 1397, 1314, 1220, 703 cm⁻ ¹.

δ [(CD₃)₂ SO]: 1.16 (3H.t.OCH₂ CH₃), 1.43 (3H.s. gem methyl), 1.54 (3H.s. gem methyl), 1.5-2.2 (4H.m. -- CH₂ CH₂ --), 4.1(1H.s.C3 proton), 4.88 (1H.m. CH CH₂ CH₂), 5.3-5.8 (3H.m β-lactams and PhCH<), 6.3 (1H.m.--NHCO*--), 7.3-7.7 (6H.m. aromatic protons and --NHCO*--), 8.2-9.0 (5H.m. 2 × --CONH*₂ ) and --CONH*

NH₂ OH Assay: 100%. Biochromatogram: Single zone R_(f) 0.34

EXAMPLE 32. D-α-[D,L-β-Phenyl-α-ureidopropionamido]-1,4-cyclohexadienylacetamidopenicillanic acid. ##SPC17##

Prepared by method Bv from D,L-β-phenyl-α-ureidopropionic acid.

Yield: 65%. νmax (KBr): 3340, 1762, 1720, 1650, 1530, 1230 and 703 cm⁻ ¹. δ [(CD₃)₂ SO]: 1.5(6H.d. gem dimethyls); 2.6(4H.s. cyclohexadiene methylenes); 2.6-3.2(2H.m. PhCH₂ CH<); 4.3(1H.s.C-3proton); 4.3-4.8(1H.m. PhCH₂ CH<);4.9-5.9 ##SPC18##

5.67(3h.s. cyclohexadiene methines);6.3-6.7(1H.m. --NHCONH₂ *);7.25(5H.s.PhCH₂ CH<);8.0-8.3(1H.m.--CONH--*);

Hydroxylamine Assay: 59%. Biochromatography: R_(f) 0.56.

EXAMPLE 33 Sodium D-β-[D-β-Phenyl-β-ureidopropionamido]-1,4-cyclohexadienylacetamido penicillanate. ##SPC19##

Prepared by method Bvi) from D-β-phenyl-α-ureidopropionic acid and epicillin.

Yield: 45%. γmax(KBr): 3350, 1760, 1630, 1530, 1230 and 703 cm⁻ ¹. δ [(CD₃)₂ SO]: 1.53(6H.d. gem dimethyls); 2.65(4H.s.cyclohexadienyl methylenes); 2.8-3.0(2H.m.PhCH₂ --); 4.0(1H.s.C-3 proton); 4.2-4.7(1H.m.)CH₂ CH--); 5.0-5.6(5H.m.β-lactam protons HNCHCO and HNCONH₂):5.7(3H.s.cyclohexadienyl vinylic protons); 6.4-6.7(1H.m.--NHCONH₂); 7.25(5H.s.Ph); 8.0-9.0(2H.m.NH) Hydroxylamine Assay: 76%. Biochromatography: 1 zone at R_(f) 0.47

EXAMPLE 34 D-α-[DL-β-Benzyloxy-α-ureidopropionamido]-phenylacetamido-penicillanic acid.

(R=Ph; R¹ =PhCH₂ OCH₂ ; R³ =H; R² =NHCONH₂ ; M=H; α¹ =DL)

Prepared by method Bi from β-benzyloxy-α-ureido-DL-propionic acid and ampicillin.

Yield: 56%.

νmax (KBr): 3350, 1770, 1650, 1520, 1220 and 700 cm⁻ ¹ δ [(CD₃)₂ SO]: 1.5(6H.d. gem dimethyls); 3.65(2H.m. --OCH₂ CHC<) 4.25(1H.s.C-3 proton); 4.5(3H.m. --OCH₂ CH< and PhCH₂ O--); 5.4-7.0(6H.m. β-lactam protons, HN-CHCO and --NHCONH₂); 7.2-7.5(10H.m. 2 × phenyl aromatics); 8.5-9.2(2H.m. amide NH's)

Analysis: C₂₂ H₃₁ N₅ O₇ S.H₂ O required: C, 55.3; H, 5.63; N, 11.9. Found : C, 56.03; H, 5.72: N, 11.46. Hydroxylamine Assay: 95%. Biochromatography: 1 zone at R_(f) 0.42.

EXAMPLE 35 D-α-[DL-β-Benzyloxy-α-ureidopropionamido]-p-hydroxyphenylacetamido penicillanic acid.

(R = p-HO--Ph; R¹ = PhCH₂ OCH₂ ; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = DL)

Prepared by method Biii from DL-β-benzyloxy-α-ureidopropionic acid and amoxycillin.

Yield: 59%. νmax (KBr): 3350, 1775, 1725, 1650, 1515, 1230, and 703 cm⁻ ¹ δ [(CD₃)₂ SO]: 1.5(6H.d. gem dimethyls); 3.5-3.9(2H.m. --OCH₂ CH<); 4.23 (1H.s. C-3 proton); 4.3-4.7(3H.m. PhCH₂ O and OCH₂ CH<); 5.2-5.9(5H.m. β-lactams, --HNCHCO and CONH₂) 6.2-6.5 (1H.m. --NHCONH₂); 6.5-7.5(9H.m. aromatic protons) 8.3-9.0 (2H.m. amide NH's)

Hydroxylamine Assay: 84%. Biochromatography: 1 zone at R_(f) 0.3.

Example 36 D-α-[D-β-Phenyl-α-ureidopropionamido]-3-thienylacetamido penicillanic acid. ##SPC20##

Prepared by method Biv from D-β-phenyl-α-ureidopropionic acid.

Yield: 59%. M.p. 175°-7°C (decomp.) νmax (KBr): 3360, 1750, 1650, 1525 and 704 cm⁻ ¹ δ [(CD₃)₂ SO]: 1.53(6H.m. gem dimethyls); 2.92(2H.m. PhCH₂ CH<) 4.28(1H.s. C-3 proton); 4.61(1H.m. PhCH₂ CH<); 5.31-6.05 (5H.m. β-lactams, CONH₂ * and ThCH<); 6.26(1H.d. CONH--*); 7.37(8H.m. Ph and Th); 8.58 and 9.07 (2 × 1H.d. --CONH--*)

Hydroxylamine Assay: 100%. Biochromatogram: 1 zone at R_(f) 0.40

Example 37 D-β-[D-α-Ureidopropionamido]-phenylacetamido penicillanic acid.

(R = Ph; R¹ = CH₃ ; R³ = H; R² = NHCONH₂ ; M = H; α¹ = D)

Prepared by method Bi from D-α-ureidopropionic acid.

Yield: 38%. M.p. 176°-8°C (decomp.) νmax(KBr): 3350 (br), 1773, 1720, 1635, 1530, 1234, and 700 cm⁻ ¹. δ [(CD₃)₂ SO]: 1.21 (3H.d. CH₃ CHNH--); 1.43 (3H.s. gem dimethyls); 1.57 (3H.s. gem dimethyls); 4.26 (1H.s. C-3 proton); 4.33(1H.m. CH₃ CHNH--); 5.33-5.91(5H.m. β-lactams, PhCH< and CONH₂ *); 5.91-6.54 (1H.m. --NHCONH₂); 7.38(5H.m. aromatic protons); 8.47 and 9.05 (2 × 1H.d. --CONH--*)

Hydroxylamine Assay: 106%. Biochromatogram: 1 zone at R_(f) 0.21.

EXAMPLE 38 D-α-[D,L-β-Phenyl-α-ureido-propionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; M = H; α¹ = D,L)

Prepared by method B from D,L-β-Phenyl-α-ureido propionic acid.

Yield: 37% νmax (KBr): 3350 (br.), 1775, 1650, 1525, 1225, 702 cm⁻ ¹. δ [(CD₃)₂ SO]: 1.45 (3H.s. gem methyl), 1.57 (3H.s. gem methyl), 2.88 (2H.m. PhCH₂ --), 4.26 (1H.s. C3 proton), 4.60 (1H.m. PhCH₂ CH<), 5.33-5.94(5H.m. β-lactams, PhCH, --CONH₂ *), 6.24 (1H.d. --CONH*--), 7.27 (10H.m. aromatic protons), 8.54 (1H.d. --CONH*--), 9.11 (1H.d. --CONH*--)

NH₂ OH Assay: 87% Biochromatogram: Single zone R_(f) 0.42.

EXAMPLE 39 D-α-[-α-Methyl-α-ureidopropionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = R³ = CH₃ --; R² = NH₂ CONH--; M = H)

Prepared by method B from α-Ureido-isobutyric acid

Yield: 6% νmax (KBr): 3400 (br.), 1785, 1715, 1650, 1535, 1225 and 700Cm⁻ ¹ δ[(CD₃)₂ SO]: 1.38 (6H.s. (CH₃)₂ C<), 1.46 (3H.s. gem methyl) 1.57 (3H.s. gem methyl), 4.27 (1H.s. C3 proton), 5.37-5.88 (5H.m. β-lactams, CONH₂ *, Ph CH<), 6.37 (1H.s. --NHCO*--), 7.40 (5H.m. aromatic protons), 8.13 and 9.08 (2 × 1H.d. --CONH*--)

NH₂ OH Assay: Biochromatogram: Single zone R_(f) 0.27. Analysis: Found: C, 52.60; H, 5.89; N, 14.34; S, 6.88% C₂₁ H₂₇ N₅ O₆ S required C, 52.82; H, 5.70; N, 14.67; S, 6.71%

EXAMPLE 40 D-α-[D,L-α-Methyl-β-phenyl-α-ureidopropionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ --; R³ = CH₃, R² = NHCONH₂, α = D,L)

Prepared by method B from D,L-α-Methyl-β-phenyl-α-ureidopropionic acid.

Yield: 43%. νmax (KBr): 3370 (br.), 1775, 1720, 1655, 1525, 1220 and 704 cm⁻ ¹. δ[(CD₃)₂ SO]: ##EQU5## 1.43 (3H.s. gem. methyl), 1.57 (3H.s. gem methyl), 2.80-3.70 (2H.m. PhCH₂ --), 4.30 (1H.m. C3 proton), 5.38-6.12 (5H.m. β-lactams, PhCH and CONH₂ *), 6.23 (1H.s. --CONH--), 7.39 (10H.m. aromatic protons), 8.34 (1H.d. --CONH--*), 9.28 (1H.d. --CONH--*)

Biochromatogram: Single zone R_(f) 0.54. Analysis: Found: C, 56,49; H, 5.66; N, 12.16% C₂₇ H₃₁ N₅ O₆ S requires C, 58.58; H, 5.64; N, 12.65%.

EXAMPLE 41 D-α-[D,L-α-Acetamido-β-phenylpropionamido]-p-hydroxyphenylacetamido penicillanic acid

(R = p HO-Ph; R¹ = PhCH₂ ; R³ = H; R² = --NHCOCH₃ ; M = H; α¹ = D,L)

Prepared by method Bii from N-Acetyl-D,L-β-phenylalanine.

Yield: 40% νmax (Nujol): 3250 (br.), 1760, 1630, 1515, 1380, 1220, 710cm⁻ ¹. δ[(CD₃)₂ SO]: 1.43 (3H.s. gem methyl), 1.57 (3H.s. gem methyl) 1.75 (3H.s. --NHCOCH₃, 3.0 (2H.m. PhCH₂), 4.24 (1H.s. C3 proton), 6.72 (1H.m. PhCH₂ CH<), 5.4-5.9 (3H.m. β-lactams and PhCH<), 6.58-7.50 (9H.m. aromatic protons), 8.10, 8.47 and 8.95 (3 × 1H.m. --CONH--*).

NH₂ OH Assay: 105% Biochromatogram: Single zone R_(f) 0.51

Example 42 D-α-[L-β-(p-Methoxyphenyl)-α-ureidopropionamide]phenylacetamido penicillanic acid (R = Ph; R¹ = pMeO-PhCH₂ --; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = L)

Prepared by method B from α Ureido-L-β-(p Methoxyphenyl) propionic acid.

Yield: 19%. νmax (KBr): 3300 (br.), 1775, 1640, 1515, 1250 and 700cm⁻ ¹. δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl), 1.58 (3H.s. gem methyl), 2.83 (2H.m. PhCH₂), 3.72 (3H.s. CH₃ O), 4,28 (1H.s. C3 proton), 4.61 (1H.m. PhCH₂ CH), 5.35-5.93 (5H.m. β-lactams, PhCH, --NHCONH₂ *), 6.27 (1H.d. --NHCO--), 6.67-7.50 (9H.m. aromatic protons), 8.70 and 9.32 (2 × 1H.d. --NHCO--*)

NH₂ OH Assay: 81% Biochromatogram: Single zone R_(f) 0.43.

EXAMPLE 43 D-α-[D-α-Guanidino-β-phenylpropionamido]-phenylacetamido penicillanic acid hydrochloride

(R = Ph; R¹ = PhCH₂ --, R³ = H; ##EQU6## M = H α¹ = D)

Prepared by method E from D-α-Guanidino-β-phenyl propionic acid.

Yield: 21% νmax (KBr): 3330 (br.), 1768, 1663, 1602, 1525, 1458 1394, 1320, 703cm⁻ ¹. δ[(CD₃)₂ SO]: 1.44 (3H.s. gem methyl), 1.57 (3H.s. gem methyl), 2.96-3.23 (2H.m. PhCH₂ --), 4.17 (1H.m. C3 proton), 4.61 (1H.m. PhCH₂ CH<), 5.30-5.94 (3H.m. β-lactams and PhCH<), 7.11-7.71 (14H.m. aromatic protons and ##EQU7## ##EQU8## 8.82 and 9.12 (2 × 1H.m. --CONH--*)

NH₂ OH Assay: 90%. Biochromatogram: Single zone R_(f) 0.52.

EXAMPLE 44 D-α-[D,L-β-Methoxy-α-ureidopropionamido]-phenyl-acetamido penicillanic acid

(R = Ph; R¹ = CH₃ OCH₂ ; R³ = H; R² = --NHCONH₂ ; M = H; α = D,L)

Prepared by method B from D-α-Ureido-β-methoxy propionic acid.

Yield: 23%. νmax (KBr): 3350 (br.), 1775, 1650, 1520, 1310, 1225, 1115 and 700cm⁻ ¹. δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl), 1.58 (3H.s. gem methyl), 3.27 (3H.d. CH₃ OCH₂ --), 3.57 (2H.m. CH₃ OCH₂ --), 4.18-4.75 (1H.m. --CH₂ CH<), 4.27 (1H.s. C3 proton), 5.23-5.98 (5H.m. β-lactams, --NHCONH₂ *, PhCH<), 6.37 (1H.d. --NHCO--*), 7.38 (5H.m. aromatic protons) 8.42 (1H.d. --CONH--*), 9.14 (1H.m. --CONH--*).

NH₂ OH Assay: 98%. Biochromatogram: Single zone R_(f) = 0.24.

EXAMPLE 45 Phthalid-3-yl D-α-[D-α-Guanidino-β-Phenylpropionamido] phenylacetamido penicillinate, hydrochloride ##SPC21##

Prepared by method A from D-α-Guanidino-β-phenyl propionic acid.

Yield: 23% νmax (KBr): 3340 (br.), 1785, 1660, 1510, 1285, 980 755 and 705cm⁻ ¹. δ[(CD₃)₂ SO]: 1.53 (6H.m. gemdimethyls), 3.03 (2.H.m. PhCH₂ CH), 4.56 (1H.s. C3 proton), 4.37-5.08 (1H.m. PhCH₂ CHC), 5.33-6.01 (3H.m. β-lactams and PhCH), 7.52 (1H.s. Phthalide 3 proton), 6.95-8.05 (19H.m. aromatic protons and ##EQU9## 8.05-9.49 (2 × 1H.m. --CONH--*)

NH₂ OH Assay: 91%. Biochromatogram: Single zone R_(f) 0.87.

EXAMPLE 46 D-α-[L-β-Phenyl-α-ureidopropionamido]-phenylacetamido penicillanic acid

(R = Ph; R¹ = PhCH₂ ; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = L)

Prepared by method Bi from D-α-Ureido-β-phenyl propionic acid.

Yield: 15% νmax (KBr): 3360 (Br.), 1775, 1650, 1525, 1315, 1230 and 705cm⁻ ¹. δ[(CD₃)₂ SO]: 1.44 (3H.s. gem methyl), 1.59 (3H.s. gem methyl), 2.89 (2H.m. PhCH₂ --), 4.25 (1H.s. C3 proton), 4.68 (1H.m. PhCH₂ CH), 5.64 (5H.m. β-lactams NHCONH₂ *), PhCH)), 6.27 (1H.d. --NHCO--), 7.28 (10H.m. aromatic protons), 8.62 and 9.17 (2 × 1H.d. --CONH--*)

Biochromatogram: Single zone

EXAMPLE 47 D-α-[D,L-α-(3-ethylureido)-β-phenylpropionamido]phenylacetamidopenicillanic acid ##SPC22##

Prepared by method Bi from D,L-(3-ethylureido)-β-phenylpropionic acid.

Yield: 16% m.p. 174°-6°C (dec.) νmax(KBr): 3380 (broad), 1774, 1637, 1540, 1299, 1218 and 701cm⁻ ¹. δ[(CD₃)₂ SO]: 0.96 (3H.t. --NHCH₂ CH₂), 1.44 (3H.s. gem methyl), 1.58 (3H.s. gem methyl), 2.96 (4H.m. PhCH₂ CH<, --NHCH₂ CH₃), 4.27 (1H.s. C3 proton), 4.59 (1H.m. PhCH₂ CH<), 5.38-6.17 (5H.m. β-lactams, PhCH<, --NHCONH--*), 7.32 (10H.m. PhCH<, PhCH₂ CH<), 8.58 (1H.m. --CONH--*), 9.12 (1H.m. --CONH--*).

Hydroxylamine Assay: 92.9%. Biochromatography: R_(f) = 0.71.

EXAMPLE 48 D-α-[D,L-β-(2-Thienyl)-α-ureidopropionamido]-phenylacetamidopenicillanic acid ##SPC23##

Prepared by method Bi) from D,L-β-(2-thienyl)-α-ureidopropionic acid.

Yield: 25.9% νmax(KBr): 3355 (br.), 1773, 1648, 1537, 1307, 1226 and 701cm⁻ ¹. δ[(CD₃)₂ SO]: 1.42 (3H.s. gem methyl), 1.56 (3H.s. gem methyl), 3.10 (2H.m. --CH₂ CH<), 4.21 (1H.s. C3 proton), 4.5 (1H.m. --CH₂ CH<), 5.58 (5H.m. β-lactams, --NHCONH₂ *, PhCH), 6.28 (1H.m. --NHCONH₂ *), 6.89 (2H.m. thienyl 3- and 4-protons), 7.35 (6H.s. phenyl aromatics and thienyl 5-proton), 8.67 (1H.m. --CONH--*), 9.15 (1H.m. --CONH--*).

Hydroxylamine Assay: 70.6%. Biochromatography: R_(f) = 0.41.

EXAMPLE 49 Potassium D-α-[D-α-(3-ethylureido)-p-phenyl-propionamido]-phenylacetamidopenicilianate (R = Ph--, R¹ = PhCH₂ --, R³ = H--, R² = --NHCONHCH₂ CH₃, N = K, α¹ = D)

Prepared by method Bi from D-α-(3-ethylureido)-β-phenylpropionic acid.

Yield: 65.3% νmax(KBr): 3350(br), 1774, 1630, 1540, 1225, 732 and 702cm⁻ ¹. δ[(CD₃)₂ SO]: 0.95 (3H.t. J = 7Hz --NHCH₂ CH₃), 1.45 (3H.s. gem-methyl), 1.58 (3H.s. gem-methyl), 2.75-3.35 (4H.m. PhCH₂ CH, --NHCH₂ CH₃ (J = 7Hz)), 4.28 (1H.s. C-3 proton), 4.61 (1H.m. PhCH₂ CH<), 5.4-6.3 (5H.m. β-lactams, PhCH, --NHCONH--*), 7.37 (10H.m. PhCH, PhCH₂ CH<), 8.53 (1H.d. --CONH--*), 9.12 (1H.m. --CONH--*),

Hydroxylamine Assay: 85.4%. Biochromatography: R_(f) = 0.65.

EXAMPLE 50 D-α-[D-β-Phenyl-α-(3-n-propylureido)-propionamido]-phenylacetamidopenicillanic acid ##SPC24##

Prepared by method Bi from D-α-(3-n-propylureido)-propionic acid.

Yield: 70.3% νmax(KBr): 3320 (br.), 1772, 1633, 1540, 1222, 731 and 701cm⁻ ¹. δ[(CD₃)₂ SO]: 0.81 (3H.t. --NHCH₂ CH₂ CH₃), 1.45 (3H.s gem methyl), 1.58 (3H.s. gem methyl), 2.7-3.3 (6H.m. PhCH₂ CH<, --NHCH₂ CH₂ CH₃), 4.27 (1H.s. C-3 proton), 4.6 (1H.m. PhCH₂ CH<), 5.4-6.3 (5H.m. β-lactams, PhCH<, --NHCONH--*), 7.31 (10H.m. PhCH<, PhCH₂ CH<), 8.53 (1H.m. --CONH--*), 9.10 (1H.m. --CONH--*).

Hydroxylamine Assay: 94.7% Biochromatography: R_(f) = 0.73

EXAMPLE 51 D-α-[D-β-Phenyl-α-(3-iso-propylureido)-propionamido]-phenylacetamidopenicillanic acid ##SPC25##

Prepared by method Bi from D-α-phenyl-α-(3-iso-propylureido-propionic acid.

Yield: 60% νmax (KBr): 3363 (br), 1772, 1626, 1533, 1230, 1128, 729 and 701 cm⁻ ¹. δ[(CD₃)₂ SO]: 0.99 (6H.d. NHCH(CH₃)₂), 1.45 (3H.s. gem methyl), 1.58 (3H.s. gem methyl), 2.92 (2H.m. PhCH₂ CH<), 3.3. (1H.m. --NHCH(CH₃)₂), 4.28 (1H.s. C-3 proton), 4.6 (1H.m. PhCH₂ CH<), 5.4-6.3 (5H.m. β-lactams, PhCH<, --NHCONH--*), 7.23 (5H.s. PhCH₂ CH<), 7.39 (5H.m. PhCH<), 8.4-9.5 (2H.m. 2 × --CONH--*).

Hydroxylamine Assay: 89.7%

Biochromatography: R_(f) = 0.72

EXAMPLE 52 Potassium D-α-[D-α-(3-Cyclohexylureido)-βphenylpropionamido]-phenylacetamidopenicillanate ##SPC26##

Prepared by method Bi from D-α-(3-cyclohexylureido)-β-phenylpropionic acid.

Yield: 57% νmax (KBr): 3330 (br.), 1762, 1628, 1546, 1392, 1320 and 701cm⁻ ¹. δ[(CD₃)₂ SO]: 0.6-1.7 (10H.m cyclohexyl methylenes) 1.44 (3H.s. gem methyl), 1.54 (3H.s. gem methyl), 2.75-3.4 (3H.m. PhCH₂ CH<, ##EQU10## 3.92 (1H.d. C-3 proton), 4.3-4.7 (1H.m. PhCH₂ CH<), 5.2-5.6 (2H.m. β-lactams), 5.65-5.94 (1H.m. PhCH<), 6.1-6.5 (2H.m. --NHCONH--*), 7.23 (5H.s. PhCH₂ CH<), 7.37 (5H.m. PhCH<), 8.4-9.0 (2H.m. 2 × --CONH--*)

Hydroxylamine Assay: 85.9% Biochromatography: R_(f) = 0.73

EXAMPLE 53 Potassium D-α-[DL-α-(3-tert-butylureido)-β-phenylpropionamido] phenylacetamidopenicillanate.

(R = Ph; R¹ = PhCH₂ --; R³ = H; R² = --NHCONHC(CH₃)₃ ; M = K; α¹ = D,L)

Prepared by method Bi from D,L-α-(3-tert-butylureido)-β-phenylpropionic acid.

Yield: 40.2%. νmax (KBr): 3360(br), 1774, 1645(br), 1540(br), 1456, 1214, 733 and 702 cm⁻ ¹ δ[(CD₃)₂ SO]: 1.22 (9H.s. --NHC(CH₃)₃); 1.45(3H.s. gem dimethyl); 1.58(3H.s. gem dimethyl); 2.89(2H.m. PhCH₂ CH<) 4.23(1H.s. C-3 proton); 4.53(1H.m. PhCH₂ CH<) 5.35-6.15(5H.m. β-lactams, PhCH< and --NHCONH--*); 7.30 (10H.m. PhCH< and PhCH₂ CH<); 8.51(1H.d. --CONH--*); 9.12(1H.d. --CONH--*).

Hydroxylamine Assay: 99.8%. Biochromatography: 1 zone at Rf 0.72.

EXAMPLE 54 D-β-[DL-β-(p-Hydroxyphenyl)-α-ureidopropionamido] phenylacetamidopenicillanic acid.

R = Ph; R¹ = p--HO--PhCH₂ ; R³ = H; R² = --NHCONH₂ ; M = h; α¹ = D,L

Prepared by method B from D,L-β-(p-hydroxyphenyl)-α-ureidopropionic acid.

Yield: 80%. νmax (KBr): 3350(br), 1772, 1650, 1517, 1230, and 703 cm⁻ ¹. δ[(CD₃)₂ SO]: 1.44(3H.s. gem methyl); 1.57 (3H.s. gem methyl) ˜2.8 (2H.m. --CH₂ CH<); 4.29 (1H.s. C-3 proton) ˜4.5 (1H.m --CH₂ CH<); 5.35-5.87 (5H.m. β-lactams, PhCH< and --NHCONH₂ *); 6.27 (1H.d. --NHCONH₂ *); 6.67 (2H.d. o-protons in p--HO--Ph ring); 6.99 (2H.d. m-protons in p--HO--Ph ring); 7.30(5H. broad s. PhCH); 8.47 (1H.m. --CONH--*) 9.12 (1H.m. --CONH--*)

Hydroxylamine Assay: 79.4%. Biochromatography: 1Zone at Rf 0.27.

EXAMPLE 55 D-α-[D,L-β-(m-Hydroxyphenyl)-α-ureidopropionamido]-phenylacetamido penicillanic acid.

(R = Ph, R¹ = m--HO--PhCH₂ --, R³ = H, R² = --NHCONH₂, M = H, α¹ = D,L)

Prepared by method B from D,L-β-(m-hydroxyphenyl)-α-ureidopropionic acid.

Yield: 46%. νmax (KBr): 3360(br), 1775, 1625, 1531, 1236, 1164, and 703 cm⁻ ¹. δ[(CD₃)₂ SO]: 1.45 (3H.s. gem methyl); 1.58 (3H.s. gem methyl); ˜2.8 (2H.m. --CH₂ CH<); 4.32 (1H.s. C-3 proton) 4.55 (1H.m. --CH₂ CH<); 5.40-5.90 (5H.m. β-lactams, PhCH< and --NHCONH₂ *); 6.35 (1H.d. --NHCONH₂ *) 6.55-7.15 (4H.m. m--HO--Ph--aromatics); 7.32 (5H. broad s. PhCH<); 8.52 (1H.d. --CONH--*); 9.16 (1H.m. --CONH--*)

Hydroxylamine Assay: 93.7%. Biochromatography: 1Zone at Rf 0.39.

EXAMPLE 56 D-α-[D-β-Phenyl-α-ureidopropionamido]-(2-thienyl)-acetamidopenicillanic acid. ##SPC27##

Prepared by method Bviii with D-β-phenyl-α-ureidopropionic acid.

Yield 44.8%. νmax (Nujol): 3310(br), 1785, 1668, 1539, 1233 and 701 cm⁻ ¹. δ[(CD₃)₂ SO]: 1.47(3H.s. gem methyl); 1.60(3H.s. gem methyl; ˜3(2H.m. PhCH₂ CH<); 3.9-4.4(1H.s. PhCH₂ CH<) 4.26(1H.s. C-3 proton); 5.3-6.6 (6H.m. β-lactams, ThCH<, and --NHCONH₂ *); 7.28(8H.m. PhCH₂ CH< and ThCH<); 7.7-9.5 (2H.m. 2 × --CONH--*)

Hydroxylamine Assay: 78.2%. Biochromatography: One zone at Rf 0.62 (plus faint zone at Rf 0.27 due to starting amino penicillin.)

EXAMPLE 57 D-α-[D-β-Phenyl-α-ureidopropionamido]-valeramido-penicillanic acid.

(R = CH₃ (CH₂)₂ --, R¹ = PhCH₂ --, R³ = H, R² = )NHCONH₂, M = H, α¹ = D)

Prepared by method Bvii from D-β-phenyl-α-ureidopropionic acid.

Yield: 30%. νmax (KBr): 3340(br), 1774, 1650, 1530, 1231, and 703cm⁻ ¹ δ[(CD₃)₂ SO]: 0.92 (3H.m. CH₃ (CH₂)₂ --); 1.1-1.7 (10H.m. gemdimethyls, CH₃ (CH₂)₂ --); 2.85-3.0 (2H.m. PhCH₂ CH<); 4.3-4.7 (2H.m. CH₃ (CH₂)₂ CH<, PhCH₂ CH<); 4.34 (1H.s. C-3 proton); 5.47-5.8 (4H.m. β-lactams, --NHCONH₂ *); 6.30 (1H.d. --NHCONH₂ *); 7.27 (5H.s. aromatics); 8.12 (1H.d. --CONH--*); 8.81 (1H.m. --CONH--*)

Hydroxylamine Assay: 78%. Biochromatography: 1 zone at Rf 0.59.

EXAMPLE 58 D-α-[D-β)Phenyl-α-ureidopropionamido]-cyclopropylacetamidopenicillanic acid. ##SPC28##

Prepared by method Bvi from D-β-phenyl-α-ureidopropionic acid.

Yield: 50%. νmax (KBr): 3345(br), 1772, 1645,(br), 1527, 1230 and 703 cm⁻ ¹ δ [(CD₃)₂ SO]: 0.25-1.25H.m. cyclopropyl ring protons) 1.51 (3H.s. gem methyl); 1.63 (3H.s. gem methyl); 2.92 (2H.m. PhCH₂ --); ##SPC29##

4.3 (1h.s. PhCH₂ CH<) 5.5-5.8 (4H.m. β-lactams and --CONH₂ *); 6.18 (1H.d. --CONH--*) 7.2-7.4 (5H.m. aromatic protons); 8.18 and 8.93 (2 × 1H.d. --CONH--*)

Hydroxylamine Assay: 95.6%. Biochromatography: 1 zone at Rf 0.37

EXAMPLE 59 D-α-[D-β-Phenyl-α-ureidopropionamido]-β-phenylpropionamidopenicillanic acid

(R = R¹ = PhCH₂ --, R³ = H, R² - --NHCONH₂, M = H, α¹ = D)

Prepared by method Bix from D-p-phenyl-α-ureidopropionic acid.

Yield: 40% νmax (KBr): 3322(br), 1725, 1638, 1534, 1302, 1231, 702cm⁻ ¹ δ[(CD)₃ SO]: 1.51 (3H.s. gem-methyl), 1.65 (3H.s. gem-methyl), 3.0 (4H.m. PhCH₂, PhCH₂), 4.32 (1H.s. C-3 proton), 4.4 and 4.8 (2 × 1H.m. PhCH₂ CH<), 5.52 (2H.m. β-lactams), 6.1 (1H.m. -- CONH--), 7.1-7.4 (10H.m. aromatic protons), 8.20 and 8.78 (2 × 1H.m. --CONH*--)

NH₂ OH Assay: 87% Biochromatography: Single zone R_(f) 0.40

EXAMPLE 60 D-α-[D-α-Ureido-n-heptanamido]-phenylacetamido penicillanic acid (R = Ph; R¹ = CH₃ (CH₂)₅ ; R³ = H; R² = --NHCONH₂ ; M = H; α¹ = D)

Prepared by method Bi, from D-Ureidoheptanoic acid. Yield: 45% ν max (KBr): 3380(br), 1763, 1650, 1600, 1538, 1401, 1323, 1234, 699cm.sup.⁻¹. δ[(CD₃)₂ SO] : 0.87 (3H.m. CH₃ (CH₂)₅), 1.0-2.0 (10H.m. CH₃ (CH₂)₅), 1.47 (3H.s. gem-methyl), 1.56 (3H.s. gem-methyl), 3.97 (1.H.s. C-3 proton), 3.4-3.8 (1H.m (CH₂)₅ CH<), 5.3-5.9 (5H.m β-lactams, PhCH<,--CONH₂ *), 7.2-7.6 (5H.m. aromaticprotons), 6.6, 8.7 and 8.9 (3 × 1H.m. --CONH* --)

NH₂ OH Assay : 37%. Biochromatography: Single zone R_(f) 0.67

EXAMPLE 61 D-α-[D-α-Ureido-n-hexanamido]-phenylacetamido penicillanic acid (R = Ph, R¹ = CH₃ (CH₂)₃, R³ = H, R² = --NHCONH₂, M = H, α¹ = D

Prepared by method Bi using D-α-ureidohexanoic acid.

Yield: 60% νmax (KBr) : 3340, 1772, 1640, 1312, 1234, 700cm.sup.⁻¹. δ[(CD₃)₂ SO] : 0.84 (3H.m CH₃ (CH₂)₃, 1.0-1.8 (6H.m. CH₃ (CH₂)₃, 1.42 (3H.s. gem-methyl), 1.54 (3H.S gem-methyl), 4.23 (1H.s. C-3 proton), 4.23 (1H.m. CH₃ (CH₂)₃ CH<), 5.4-5.9 (5H.m. β-lactams, PhCH< and CONH₂ *), 6.25 (1H.m. --CONH*--), 7.40 (5H.m. aromatic protons), 8.49 and 9.08 (2 × 1H.d. --CONH--)

Hydroxylamine Assay: 73%

Biochromatography: R_(f) = 0.59

EXAMPLE 62 D-α-[D-β-(1,4-Cyclohexadienyl-α-ureidopropionamido]-(p-hydroxyphenyl)acetamidopenicillanic acid ##SPC30##

Prepared by method Bii) using D-β-(1,4-cyclohexadienyl)-α-ureidopropionic acid

Yield: 50% νmax (KBr): 3330(br), 1770, 1640, 1510, 1223, 961, and 840cm.sup.⁻¹. δ[(CD₃)₂ SO] : 1.45 (3H.s. gem-methyl); 1.58 (3H.s. gem-methyl); 2.1-2.4 ##SPC31##

2.61 (4H. broad s. cyclohexadiene methylenes); 4.24 ##SPC32##

5.35-5.8 ##SPC33##

5.9-6.2 (1H.m. --NHCONH₂ *); 6.72 ##SPC34## ##SPC35##

8.3 (1h.m. --CONH--*); 8.9 (1H.m. H.m. --CONH*). * removable with D₂ O

Hydroxylamine Assay: 98.2% Biochromatography: R_(f) = 0.41

EXAMPLE 63

D-α-[D-β-(1,4-Cyclohexadienyl)-α-(3-ethylureido)-propionamido]-(p-hydroxyphenyl)-acetamido-penicillanic acid ##SPC36##

Prepared by method Bii from D-β-(1,4-cyclohexadienyl)-α-(3-ethylureido)-propionic acid.

Yield: 20%

νmax (KBr): 3350 (br), 1760, 1510, 1371, 1258, 1220 and 781cm.sup.⁻¹. δ[(CD₃)₂ SO]: 1.1-1.7 (9H.m. gem dimethyls, --NHCH₂ CH₃) 1.9-2.2 ##SPC37##

2.45-2.75 (4h.m. cyclohexadiene methylenes), 3.9-4.5 ##SPC38##

6.6-7.6 (4H.m aromatics), 8.3-9.2 (2H.m 2 × --CONH *--).

Hydroxylamine Assay: 88% Biochromatography: R_(f) = 0.44 (plus 2 minor zones)

EXAMPLE 64 D-α-[D-α-Acetamido-n-hexanamido]-(p-hydroxyphenyl)-acetamidopenicillanic acid ##SPC39##

Prepared by method Bii from D-α-acetamido-n-hexanoic acid.

Yield: 30% νmax (KBr): 3310(br.), 1770, 1645, 1510, 1374, 1210, 1173cm.sup.⁻¹. δ[(CD₃)₂)SO]: 0.8-1.8 (9H.m. CH₃ (CH₂)₃ --), 1.43 (3H.s. gem methyl), 1.54 (3H.s. gem methyl), 1.92 (3H.d. COCH₃) 4.28 (1H.s. C3 proton), 4.38 (1H.m CH₂ CH<), 5.4-5.8 (3H.m. β-lactams and PhCH<), 6.22 (1H.d. --CONH*--), 7.0-7.6 (4H.m. aromatic protons), 8.25 and 9.3 (2 × 1H.m. --CONH--*)

Hydroxylamine Assay: 124.6% Biochromatography: R_(f) = 0.35 (plus small zone due to amino-penicillin). 

We claim:
 1. A penicillin of formula (I): ##SPC40##wherein R is phenyl, hydroxyphenyl, halophenyl, nitrophenyl, alkoxyphenyl having 1-3 carbon atoms in the alkoxy part, aminophenyl, 2- or 3-thienyl, cycloalkyl having 3-7 carbon atoms in the cyclo part, cycloalkenyl having 5-7 carbon atoms in the cyclo part, or alkyl having 1-4 carbon atoms; R¹ is cyclohexa-1,4-dienyl, cyclopropyl or cyclohexyl; R² is ##EQU11## in which R⁵ is amino, mono- or di-alkylamino having 1-4 carbon atoms in the alkyl part, cyclohexylamino, hydrogen, alkyl of 1-4 carbon atoms or phenyl, and R⁶ is amino, mono- or di-alkylamino having 1-4 carbon atoms in the alkyl part or cyclohexylamino; R³ is hydrogen or alkyl of 1-3 carbon atoms; or a pharmaceutically acceptable salt or a pharmaceutically acceptable hydrolyzable ester which converts to the free acid from in vivo.
 2. A penicillin of claim 1 wherein the carbon atom attached to R is in the D-configuration.
 3. A penicillin of claim 1 wherein the carbon atom attached to R² is in the D-configuration.
 4. A penicillin of claim 1 wherein R is phenyl, 4-hydroxyphenyl or 3-thienyl.
 5. A penicillin of claim 1 wherein R³ is hydrogen.
 6. A penicillin of claim 1 wherein, when R² is (II), R⁵ is amino or hydrogen or, when R² is (III), R⁶ is amino.
 7. A penicillin of claim 1 wherein the ester is phthalidyl, 5,6-dimethoxyphthalidyl, pivaloyloxymethyl or acetoxymethyl.
 8. A penicillin of claim 1 wherein the salt is a base addition or acid addition salt. 